Objective To explore the pathological features affecting the prognosis by observing lung adenosquamous carcinoma overall survival after surgical treatment.Methods Totally 59 cases of lung ASC from 2531 surgically treated lung cancer patients in the First Hospital of Jilin University,from January 2000 to June 2012,were retrospectively analyzed to study their clinical characteristics,survival condition and the related factors influencing the prognosis.Using log-rank test and Cox multiple factors analysis for statistical analysis.Results (1) The 59 patients with ASC were mostly the male patients (62.7％).The median age was 57.2 years.Median survival time was 409 days(13.6 months).1-,3-,5-year survival rates were 59.9 ％,36.4％ and 31.2 ％.(2) Among the 59 patients (52 cases of pathological specimens),11 cases were EGFR mutation positive,positive rate was 21.2％,2 cases of patients were KRAS mutations positive,positive rate was 3.8％ ;(3) Single factor and multiple factors analysis showed that the pathological subtype,adjuvant treatment,pleural invasion and tumor stage were associated with prognosis as independent factors (P ＜ 0.05).Conclusion Compared with the simplex lung squamous carcinoma and lung adenocarcinoma,lung adenosquamous carcinoma has poorer prognosis.Early diagnosis and given comprehensive treatment were the keys to prolong its survival.

Objective To study postoperative delirium in elderly patients.Methods We investigate the morbidity of postoperative delirium in 142 elderly patients (≥ 60 years)after gastrointestinal surgery by using Confusion Assessment Method (CAM) and Delirium Rating Scale Revised-98 (DRS-R98) scores.Data were analyzed using Student's t test and Chi-squaretest respectively with SPSS 19.0.Results Of 142 patients,delirium was diagnosed in 36 patients(25.4％),delirium developed in 4,7,17,7,1 patients in posto perative 1,2,3,4-7,7 + days respectively.There were significant difference in hospital stay:17.7 ± 2.6 days (postoperative delirium) and 13.4 ± 2.3 days (no postoperative delirium),t =4.608,P =0.000 1.The postoperative complications (52.8％ / 23.6％,x2 =10.710,P =0.001) and ICU admission (22.2％/6.6％,x2 =6.939,P =0.008) significantly increased.Conclusions Postoperative delirium is recognized as one of the most common surgical complications in elderly patients with gastrointestinal surgery leading to other major postoperative complications,and prolonged hospitalization.

BACKGROUNDThe vertebral artery (VA) and atlantoaxial joint (AAJ), with complicated structures, are located in the depths of the head-neck boundary area, the regional anatomy of which cannot be shown globally and directly. This study aims to evaluate three-dimensional CT angiography (3DCTA) in displaying the AAJ, atlantoaxial segment of the vertebral artery (ASVA) and the identification of their interrelations.

METHODSSixty-eight subjects without pathology of the ASVA and AAJ were selected from head-neck CTA examination. All the 3D images were formed with volume rendering (VR) together with techniques of separating, fusing, opacifying and false-coloring (SFOF). On the 3D images, the ASVA and AAJ were observed, and their interrelations were measured.

RESULTSAll the 3DCTA images were of high quality and up to our requirements. They could clearly and directly show the ASVA, ascending along the AAJ. There were 5 curves in the course of the ASVA, of which 2 curves were away from the atlantoaxial joint, one in the 2nd curve of 0.0 mm - 5.4 mm, the other in the 4th of 2.6 mm - 9.2 mm. There was no significant difference in the measurements between left and right (P > 0.05). The curved parts of the ASVA slightly expanded, with the biggest diameter of 5.6 mm in the 4th curve. Statistical comparison shows that the left ASVA is larger than the right (P < 0.05). Variations of the ASVA were found in 8 cases and of the AAJ in 12.

CONCLUSIONS3DCTA can globally and directly demonstrate the structures of the AAJ, ASVA and their interrelations. The 3D imaging data make up and enrich the research contents of regional anatomy and lay the foundation for related study and applications.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Atlanto-Axial Joint ; diagnostic imaging ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Vertebral Artery ; diagnostic imaging ; Young Adult

Objective To explore the effect of recovery sleep on the executive function after 36 h of total sleep deprivation by event related potential technology.Methods Thirteen healthy male college students participated in two trials. At the first trial normal sleep as control was investigated. At the second trial participants experienced 36 h of sleep deprivation and then accepted 8 h recovery sleep. In each trial six Go/Nogo tests were employed to test the executive control function and the ERP data were recorded. Results There was no statistical difference in behavior and ERP results at each time point as the subjects had normal sleep. After 36 h of sleep deprivation, the behavior results were statistically significant when compared to the baseline. The amplitude and latency of Nogo-N2, Nogo-P3 on Fz electrode, the amplitude and latency of Nogo-P3 on Cz electrode showed statistical significance when compared to the baseline. After 8 h recovery sleep, the average correct reaction time and the Go correct reaction rate had statistical significance compared to 36 h value. The amplitude of Nogo-N2 and Nogo-P3 had no statistical significance compared to the baseline.However,it was of statistical significance[(-6.80 3.95)vs(-3.37 2.63)μV,(10.63±6.62)vs(5.63±5.45)μV,(9.49±7.37)vs(6.08±6.56)μV] compared to 36 h value. The latency of the recovery value of Nogo-N2 and Nogo-P3 was statistically significant[(254.14±15.55)vs(243.08±13.97)ms(382.14±41.07)vs(349.17±30.36)ms,(369.86±26.48)vs(347.48±29.24)ms]compared to the baseline.Conclusion As the time of sleep deprivation is prolonged, the executive function is impaired and the executive function is not completely recovered after 8 h recovery sleep.

Objective To clone,express and characterize a tegument protein gene of Schistosoma japonicum(Sj29),and investigate the immune protection of the recombinant protein against S.japonicum in mice.Methods The gene coding for Sj29 protein was amplified by PCR,and the sequence was analyzed by bioinformatics tools.Partial fragment of Sj29 gene was subcloned into the prokaryotic expression vector pET28c(+).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced the recombinant with IPTG.The recombinant protein(rSj29) was purified by His-binding-resin affinity chromatography and characterized by Western blotting.Three groups each with 10 BALB/c mice were immunized subcutaneously three times(two weeks interval) respectively with 100 ?l recombinant rSj29(0.1 mg/ml),adjuvant or PBS.At the 15th day after the final inoculation,each mouse was challenged by 40 ?2 cercariae of S.japonicum.At the 53th day after infection,the mice were sacrificed to obtain the number of adult worms,number of eggs in liver and feces.Serum samples were collected at pre-immunization and certain time after immuniza-tion,and were analyzed for IgG by ELISA.The localization of rSj29 in worms of different developmental stages was demonstrated by immunofluorescent technique.mRNA expression level of Sj29 gene in worms of different developmental stages and three groups after infection was detected by quantitative real-time PCR.Results A 576 bp Sj29 gene fragment was obtained.The recombinant protein rSj29 with Mr 22 900 was expressed in the form of inclusion body.The recombinant rSj29 can be recognized by sera of mice immunized with rSj29 and sera of infected mice.The number of adult worms(15.4?5.9),number of hepatic eggs(40 143.3?2 995.9) and number of fecal eggs(3 803.9?110.9) in re-combinant protein group were significantly higher than those of PBS control group(20?3.4,49 318.1?6 648.3,5 238.1? 303.5,respectively)(P

The total RNA was extracted from ginseng leaves of Panax ginseng. The Cu/Zn-SOD gene was amplified via RT-PCR and the pET-28(a)-Cu/Zn-SOD expression vector was constructed. The pET-28 (a)-Cu/Zn-SOD recombinant plasmid was transformed into Escherichia coli BL21 (DE3) competent cells and was induced by IPTG in order to select optimal induction of expression conditions. The target protein was purified by the nickel ions (Ni ) affinity chromatography and the target protein enzyme activity was determinated by the xanthine oxidase method. The similarity of the Cu/Zn-SOD gene sequences and the Cu/Zn-SOD gene sequences of Korean ginseng in NCBI was 99. 00%. The target protein expression level was about 44.42%, and the molecular weight was 16.30 kDa after the pET-28(a)-Cu/Zn-SOD recombinants were induced by IPTG. The purified Cu/Zn-SOD protease activity reached 10,596.69 U x mg(-1). The P. ginseng pET-28(a)-Cu/Zn-SOD prokaryotic expression vector was built by the method of molecular biology, which provided the foundation for studying the Cu/Zn-SOD biology function.
Cloning, Molecular ; Escherichia coli ; genetics ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Panax ; enzymology ; genetics ; Sequence Analysis ; Superoxide Dismutase ; genetics ; isolation & purification ; metabolism

Objective Lysine-specific demethylase 1 (LSD1) is a flavin adenine dinucleotide-dependent oxidase, which participates in many biological processes , such as cell proliferation and differentiation and gene activation and repression .The aim of this study was to investigate LSD1 acetylation by histone deacetylase inhib -itor trichostatin A ( TSA) and its effect on TSA-induced apoptosis of ovarian cancer cells . Methods LSD1 shRNA was synthesized and implanted into the pLKO-Tet-On lentiviral vector , which was transfected into HO8910 and SKOV3 ovarian cancer cell lines , and then the transfected cells were screened with 1.5μg/mL puromycin for one week until stable clones were established .The cells were treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), TCP (100μmol/L), or TSA+TCP.And in the experiment of RNA interfering the LSD1 expression, the cells were also treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), Dox (100 ng/mL), or TSA +Dox.The levels of LSD1 acetylation and its substrate histone H3 lysine 4 dimethylation (H3K4me2) were de-tected by immunoprecipitation (IP) and Western blot.The apoptosis of the cells was determined by Annexin Ⅴ/PI staining and flow cytometry, the transcription levels of the Bax and p21 genes detected by real-time quantitative PCR, and the H3K4me2levels in the promoter regions of Bax and p21 measured by chromatin immunoprecipitation ( ChIP ) .Results In comparison with the methanol control, the TSA group showed significantly increased levels of LSD 1 acetylation in the HO8910(1.00±0.29 vs 5.83±0.46, P<0.01) and SKOV3 cells ( 1.00±0.24 vs 5.07±0.35, P<0.01) as well as that of H3K4me2 ( P<0.01);the total apoptosis rates of HO 8910 and SKOV3 cells were remarkably increased in the TSA, TCP, and TSA+TCP groups (P<0.05), even more significantly in the TSA+TCP than in the TSA and TCP groups ( P<0.05) .The mRNA expressions of Bax and p21 in the HO8910 cells were markedly upregulated in the TSA, Dox, and TSA+Dox groups (P<0.05), even more significantly in the latter than in the former two groups (P<0.05).The TSA group exhibited a higher level of H 3K4me2 than the methanol control in the promoters of Bax(2 .92±0.26 vs 0.68±0.19, P<0.01) and p21 (3.07±0.29 vs 0.93±0.17, P<0.01). Conclusion TSA induces the LSD1 acetylation, while suppression of LSD1 expres-sion and activity may enhance the antitumor activity of TSA .

Objective:To investigate the correlation between the location and volume of cerebral microbleeds (CMBs) and lacunar infarction (LI) in patients with cerebral small vessel disease (CSVD).Methods:Participants from the CSVD cohort in the Department of Neurology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Medical School of Nanjing University from February 2017 to March 2019 were enrolled retrospectively. All participants underwent magnetic resonance imaging scans, standardized clinical assessment and diagnosis. AccuBrain, an automatic brain segmentation and quantification software developed by the Chinese University of Hong Kong, was used to quantitatively analyze the volume of CMBs and white matter hyperintensities (WMHs). Ordered multi-class logistic regression analysis was used to determine the independent influencing factors of LI, and then multiple linear regression analysis was used to investigate the correlation between the volume of deep or infratentorial CMBs (DI-CMBs) and the number of LI. Results:A total of 317 patients with CSVD were included in the analysis, including 214 (67.5%) in the non-LI group, 43 (13.6%) in the single LI group, and 60 (18.9%) in the multiple LI group. The comparison of the three groups showed that older age, male, smoking, drinking, history of previous stroke or transient ischemic attack (TIA), lower levels of high-density lipoprotein cholesterol, larger CMBs and WMHs volume, higher enlarged perivascular spaces (EPVS) grade might be the risk factors for LI. Ordinal multivariable logistic regression analysis showed that male (odds ratio [ OR] 2.058, 95% confidence interval [ CI] 1.084-3.909; P=0.027), previous stroke or TIA history ( OR 1.962, 95% CI 1.089-3.535; P=0.025), larger WMH volume ( OR 8.716, 95% CI 4.034-18.832; P<0.001), higher EPVS grade ( OR 1.915, 95% CI 1.292-2.839; P=0.001), larger DI-CMB volume ( OR 1.022, 95% CI 1.006-1.038; P=0.008) or more DI-CMB number ( OR 1.187, 95% CI 1.005-1.403; P=0.044) were the independent related factors of LI. Multiple linear regression analysis showed that there was a significant correlation between the volume of DI-CMB and the number of LI ( r=0.330, P<0.001). Conclusion:In patients with CSVD, there is a significant correlation between DI-CMBs and LI.

Babesia microti is one of the most common causative agents of babesiosis. A sensitive and rapid detection is necessary for screening potentially infected individuals. In this study, B. microti cytochrome c oxidase subunit I (cox1) was selected as the target gene, multiple primers were designed, and optimized by a recombinase-aided amplification (RAA) assay. The optimal primers and probe were labeled with fluorescein. The sensitivity of fluorescent RAA (fRAA) was evaluated using gradient diluents of the cox1 recombinant plasmid and genomic DNA extracted from whole blood of B. microti infected mice. The specificity of fRAA was assessed by other transfusion transmitted parasites. The analytical sensitivity of the fRAA assay was 10 copies of recombinant plasmid per reaction and 10 fg/µl B. microti genomic DNA. No cross-reaction with any other blood-transmitted parasites was observed. Our results demonstrated that the fRAA assay would be rapid, sensitive, and specific for the detection of B. microti.

OBJECTIVETo observe the effects of ryanodine on rapamycin treated endothelial outgrowth cells (EOCs).

METHODSThe mononuclear cells were harvested from umbilical cord blood by Ficoll density gradient centrifugation, then induced into EOCs and expanded in vitro. The endothelial characteristics of EOCs were identified by immunostaining and fluorescent staining. The EOCs were pretreated with or without ryanodine (10 µmol/L) for 1 h, and then treated with or without rapamycin (10 nmol/L) for 24 h. Proliferation was evaluated by CCK8 and migration was measured by Transwell. The protein expression of EOCs was evaluated by immunobloting technique with total eNOS antibody and phospho-eNOS (Thr495) antibody.

RESULTSCompared with control group, the proliferation and migration capacities of EOCs were significantly reduced while the phosphorylation of eNOS (Thr495) protein was significantly upregulated in rapamycin group (P < 0.05), expression of total eNOS was not affected by rapamycin (P > 0.05). Compared with rapamycin group, the proliferation and migration capacities of EOCs were significantly increased and the phosphorylation of eNOS (Thr495) protein was significantly downregulated in ryanodine + rapamycin group (P < 0.05). The proliferation and migration capacities, the phosphorylation of eNOS (Thr495) protein and the expression of total eNOS were not affected by ryanodine alone (P > 0.05).

CONCLUSIONSRapamycin reduced proliferation and migration capacities while upregulated the phosphorylation of eNOS (Thr495) protein of EOCs and these effects could be partly reversed by cotreatment with ryanodine.

Cells, Cultured ; Down-Regulation ; Drug Synergism ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Nitric Oxide Synthase Type III ; metabolism ; Phosphorylation ; Ryanodine ; pharmacology ; Sirolimus ; pharmacology